Kinetic characterization of glucose aerodehydrogenase from Aspergillus niger EMS-150-F after optimizing the dose of mutagen for enhanced production of enzyme

In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL-1, 1118.81 s-1 and 55.94 s-1 mM-1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.

Impact of transgenic cotton varieties on activity of enzymes in their rhizosphere.

The impact of five Bacillus thuringiensis (Bt) cotton varieties and their respective isogenic non-Bt(NBt) isolines (ANKUR-2534, MECH-6304, RCH-317, ANKUR-651 and MECH-6301) was assessed on the key soil enzymes i.e., dehydrogenase, alkaline phosphatase and urease in their rhizosphere at four growth stages of the crop, namely vegetative, flowering, bolling and harvesting. These varieties were grown on farmer’s field in villages 22 miles and 24 miles of Ganganagar District of Rajasthan State in India. Results showed that dehydrogenase, alkaline phosphatase and urease activities were higher in rhizosphere of Bt isolines as compared to NBt isolines of all the varieties. Except phosphatase, differences in dehydrogenase and urease activities in rhizosphere of Bt and NBt isolines of all five varieties were significant (P<0.05). Maximum enhancement in the three enzymes activities was observed in MECH-6304 Bt isoline rhizosphere. Maximum and minimum activities of dehydrogenase and urease were observed in MECH-6304 and RCH-317 Bt isolines, respectively, whereas phosphatase activity was maximum and minimum in MECH-6304 and ANKUR-651 Bt isolines, respectively. Maximum dehydrogenase and urease activities were observed at boll formation and minimum at flowering and harvesting stage, respectively, while maximum phosphatase activity was observed at vegetative stage and minimum at harvesting stage. In conclusion, all the studied Bt isolines of cotton varieties showed no adverse effect on dehydrogenase, alkaline phosphatase and urease activities in the rhizosphere.

Inhibition of methanogenesis by interaction of aluminium ion with co-factor, F-420, in Methanosarcina barkeri.

Methane emission was inhibited by aluminium ion in paddy fields. Addition of Al3+ (20 mM) to the culture medium containing cells of pure Methanosarcina barkeri, inhibited methanogenesis. Methanogenic co-factor, F-420, was isolated and purified from Methanosarcina barkeri MS. Spectrophotometric and spectrofluorometric analysis of interaction between co-factor, F-420, and Al3+ revealed that they formed a complex compound that might have blocked methanogenesis.

Immobilization of Amaranthus leaf oxalate oxidase on alkylamine glass.

A membrane bound oxalate oxidase from leaves of Amaranthus spionsus has been partially purified and immobilized on alkylamine glass with a yield of 9.2 mg protein/g support. The enzyme retained 99.4% of initial activity of free enzyme after immobilization. There was no change in the optimum pH (3.5) and Vmax but the temperature for maximum activity was slightly decreased (35 degrees C) and energy of activation (Ea) and Km for oxalate were increased after immobilization. The immobilized enzyme preparation was stable for 6 months, when stored in distilled water at 4 degrees C. Presence of Cl- did not affect the activity of immobilized enzyme.

Noçöes de química da radiaçäo em sistemas biológicos / Notions of chemistry radiation in biological systems

A presente publicaçäo examina alguns aspectos dos efeitos biológicos diretos e indiretos da radiaçäo: formaçäo de pares, radicais livres, ion superóxido de hidrogênio, radical hidroxila, oxigênio singleto junto aos mecanismos radioprotetores endógenos dos organismos e os meios em que pode ser incrementada a radiorresistência de sistemas bioquímicos